Targeting FHL2‑E‑cadherin axis by miR‑340‑5p attenuates colon cancer cell migration and invasion

Algaber,Anwar; Madhi,Raed; Hawez,Avin; Rönnow,Carl-Fredrik; Rahman,Milladur

doi:10.3892/ol.2021.12898

Targeting FHL2‑E‑cadherin axis by miR‑340‑5p attenuates colon cancer cell migration and invasion

Authors:
- Anwar Algaber
- Raed Madhi
- Avin Hawez
- Carl-Fredrik Rönnow
- Milladur Rahman
View Affiliations

Affiliations: Department of Clinical Sciences, Malmö, Section for Surgery, Lund University, 214 28 Malmö, Sweden
Published online on: July 3, 2021 https://doi.org/10.3892/ol.2021.12898
Article Number: 637
Copyright : © Algaber et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Convincing data has suggested that four and a half LIM domain 2 protein (FHL2) serves a key function in cancer cell metastasis and that microRNA (miR)‑340‑5p can regulate cancer cell migration. The current study hypothesized that targeting FHL2 expression by miR‑340‑5p in colon cancer may attenuate colon cancer cell migration and invasion. FHL2 expression was therefore assessed in colon cancer microarray datasets using Qlucore omics explorer as well as in HT‑29 and AZ‑97 colon cancer cell lines via reverse transcription‑quantitative PCR (RT‑qPCR). Colon cancer cell migration and invasion were evaluated in the presence of miR‑340‑5p mimic, mimic control or mimic with a target site blocker. Confocal microscopy and RT‑qPCR were subsequently performed to assess FHL2, E‑cadherin (E‑cad) protein and mRNA expression in colon cancer cells. Microarray dataset analysis revealed that FHL2 expression was lower in primary colon cancer cells compared with normal colonic mucosa. It was revealed that the expression of miR‑340‑5p and FHL2 were inversely related in serum‑grown and low‑serum conditions in HT‑29 and AZ‑97 cells. Short‑time serum exposure to low‑serum grown cells induced FHL2 expression. Transfection of HT‑29 cells with miR‑340‑5p mimic not only decreased serum‑induced expression of FHL2 but also decreased cancer cell migration and invasion. Bioinformatics analysis revealed that FHL2 mRNA had one putative binding site for miR‑340‑5p at the 3'‑untranslated region. Blocking of the target site using a specific blocker reverted miR‑340‑5p mimic‑induced inhibition of FHL2 expression and cancer cell migration and invasion. Confocal microscopy confirmed that the reduction of FHL2 expression by miR‑340‑5p mimic also reversed serum‑induced E‑cad disruption and that the target site blocker abrogated the effect of miR‑340‑5p. The current results suggested that miR‑340‑5p could be used to antagonize colon cancer cell metastasis by targeting the FHL2‑E‑cad axis.

View Figures

View References

1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Miller KD, Nogueira L, Mariotto AB, Rowland JH, Yabroff KR, Alfano CM, Jemal A, Kramer JL and Siegel RL: Cancer treatment and survivorship statistics, 2019. CA Cancer J Clin. 69:363–385. 2019. View Article : Google Scholar : PubMed/NCBI
3	Bockelman C, Engelmann BE, Kaprio T, Hansen TF and Glimelius B: Risk of recurrence in patients with colon cancer stage II and III: A systematic review and meta-analysis of recent literature. Acta Oncol. 54:5–16. 2015. View Article : Google Scholar : PubMed/NCBI
4	Levin VA, Panchabhai SC, Shen L, Kornblau SM, Qiu Y and Baggerly KA: Different changes in protein and phosphoprotein levels result from serum starvation of high-grade glioma and adenocarcinoma cell lines. J Proteome Res. 9:179–191. 2010. View Article : Google Scholar : PubMed/NCBI
5	Ghosh T, Varshney A, Kumar P, Kaur M, Kumar V, Shekhar R, Devi R, Priyanka P, Khan MM and Saxena S: MicroRNA-874-mediated inhibition of the major G1/S phase cyclin, CCNE1, is lost in osteosarcomas. J Biol Chem. 292:21264–21281. 2017. View Article : Google Scholar : PubMed/NCBI
6	Rasool RU, Nayak D, Chakraborty S, Jamwal VL, Mahajan V, Katoch A, Faheem MM, Iqra Z, Amin H, Gandhi SG and Goswami A: Differential regulation of NM23-H1 under hypoxic and serum starvation conditions in metastatic cancer cells and its implication in EMT. Eur J Cell Biol. 96:164–171. 2017. View Article : Google Scholar : PubMed/NCBI
7	Huang Z, Li Q, Luo K, Zhang Q, Geng J, Zhou X, Xu Y, Qian M, Zhang JA, Ji L and Wu J: miR-340-FHL2 axis inhibits cell growth and metastasis in ovarian cancer. Cell Death Dis. 10:3722019. View Article : Google Scholar : PubMed/NCBI
8	Zhang W, Jiang B, Guo Z, Sardet C, Zou B, Lam CS, Li J, He M, Lan HY, Pang R, et al: Four-and-a-half LIM protein 2 promotes invasive potential and epithelial-mesenchymal transition in colon cancer. Carcinogenesis. 31:1220–1229. 2010. View Article : Google Scholar : PubMed/NCBI
9	Tran MK, Kurakula K, Koenis DS and de Vries CJ: Protein-protein interactions of the LIM-only protein FHL2 and functional implication of the interactions relevant in cardiovascular disease. Biochim Biophys Acta. 1863:219–228. 2016. View Article : Google Scholar : PubMed/NCBI
10	Kinoshita M, Nakagawa T, Shimizu A and Katsuoka Y: Differently regulated androgen receptor transcriptional complex in prostate cancer compared with normal prostate. Int J Urol. 12:390–397. 2005. View Article : Google Scholar : PubMed/NCBI
11	Hou Y, Wang X, Li L, Fan R, Chen J, Zhu T, Li W, Jiang Y, Mittal N, Wu W, et al: FHL2 regulates hematopoietic stem cell functions under stress conditions. Leukemia. 29:615–624. 2015. View Article : Google Scholar : PubMed/NCBI
12	Gabriel B, Fischer DC, Orlowska-Volk M, zur Hausen A, Schüle R, Müller JM and Hasenburg A: Expression of the transcriptional coregulator FHL2 in human breast cancer: A clinicopathologic study. J Soc Gynecol Investig. 13:69–75. 2006. View Article : Google Scholar : PubMed/NCBI
13	Gabriel B, Mildenberger S, Weisser CW, Metzger E, Gitsch G, Schüle R and Müller JM: Focal adhesion kinase interacts with the transcriptional coactivator FHL2 and both are overexpressed in epithelial ovarian cancer. Anticancer Res. 24:921–927. 2004.PubMed/NCBI
14	Jin H, Lee K, Kim YH, Oh HK, Maeng YI, Kim TH, Suh DS and Bae J: Scaffold protein FHL2 facilitates MDM2-mediated degradation of IER3 to regulate proliferation of cervical cancer cells. Oncogene. 35:5106–5118. 2016. View Article : Google Scholar : PubMed/NCBI
15	Bates RC and Mercurio AM: The epithelial-mesenchymal transition (EMT) and colorectal cancer progression. Cancer Biol Ther. 4:365–370. 2005. View Article : Google Scholar : PubMed/NCBI
16	Beavon IR: The E-cadherin-catenin complex in tumour metastasis: Structure, function and regulation. Eur J Cancer. 36:1607–1620. 2000. View Article : Google Scholar : PubMed/NCBI
17	Christofori G and Semb H: The role of the cell-adhesion molecule E-cadherin as a tumour-suppressor gene. Trends Biochem Sci. 24:73–76. 1999. View Article : Google Scholar : PubMed/NCBI
18	Frixen UH, Behrens J, Sachs M, Eberle G, Voss B, Warda A, Löchner D and Birchmeier W: E-cadherin-mediated cell-cell adhesion prevents invasiveness of human carcinoma cells. J Cell Biol. 113:173–185. 1991. View Article : Google Scholar : PubMed/NCBI
19	Berx G, Cleton-Jansen AM, Nollet F, de Leeuw WJ, van de Vijver M, Cornelisse C and van Roy F: E-cadherin is a tumour/invasion suppressor gene mutated in human lobular breast cancers. EMBO J. 14:6107–6115. 1995. View Article : Google Scholar : PubMed/NCBI
20	Padmanaban V, Krol I, Suhail Y, Szczerba BM, Aceto N, Bader JS and Ewald AJ: E-cadherin is required for metastasis in multiple models of breast cancer. Nature. 573:439–444. 2019. View Article : Google Scholar : PubMed/NCBI
21	Zhang W, Wang J, Zou B, Sardet C, Li J, Lam CS, Ng L, Pang R, Hung IF, Tan VP, et al: Four and a half LIM protein 2 (FHL2) negatively regulates the transcription of E-cadherin through interaction with Snail1. Eur J Cancer. 47:121–130. 2011. View Article : Google Scholar : PubMed/NCBI
22	Kreutziger KL and Kreutziger KL: Comprehensive surgical management of mandibular fractures. South Med J. 85:506–518. 1992. View Article : Google Scholar : PubMed/NCBI
23	Esteller M: Non-coding RNAs in human disease. Nat Rev Genet. 12:861–874. 2011. View Article : Google Scholar : PubMed/NCBI
24	He L and Hannon GJ: MicroRNAs: Small RNAs with a big role in gene regulation. Nat Rev Genet. 5:522–531. 2004. View Article : Google Scholar : PubMed/NCBI
25	Lim LP, Lau NC, Garrett-Engele P, Grimson A, Schelter JM, Castle J, Bartel DP, Linsley PS and Johnson JM: Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature. 433:769–773. 2005. View Article : Google Scholar : PubMed/NCBI
26	Valastyan S and Weinberg RA: Roles for microRNAs in the regulation of cell adhesion molecules. J Cell Sci. 124((Pt 7)): 999–1006. 2011. View Article : Google Scholar : PubMed/NCBI
27	Fridrichova I and Zmetakova I: MicroRNAs contribute to breast cancer invasiveness. Cells. 8:13612019. View Article : Google Scholar : PubMed/NCBI
28	Zhao P, Ma W, Hu Z, Zhang Y, Zhang S and Wang Y: Up-regulation of miR-340-5p promotes progression of thyroid cancer by inhibiting BMP4. J Endocrinol Invest. 41:1165–1172. 2018. View Article : Google Scholar : PubMed/NCBI
29	Shi Z, Li Y, Qian X, Hu Y, Liu J, Zhang S and Zhang J: miR-340 inhibits triple-negative breast cancer progression by reversing EZH2 mediated miRNAs dysregulated expressions. J Cancer. 8:3037–3048. 2017. View Article : Google Scholar : PubMed/NCBI
30	Huang D, Qiu S, Ge R, He L, Li M, Li Y and Peng Y: miR-340 suppresses glioblastoma multiforme. Oncotarget. 6:9257–9270. 2015. View Article : Google Scholar : PubMed/NCBI
31	Chen CP, Sun ZL, Lu X, Wu WX, Guo WL, Lu JJ, Han C, Huang JQ and Fang Y: miR-340 suppresses cell migration and invasion by targeting MYO10 in breast cancer. Oncol Rep. 35:709–716. 2016. View Article : Google Scholar : PubMed/NCBI
32	Matsushita Y, Hoff SD, Nudelman ED, Otaka M, Hakomori S, Ota DM, Cleary KR and Irimura T: Metastatic behavior and cell surface properties of HT-29 human colon carcinoma variant cells selected for their differential expression of sialyl-dimeric Le(x)-antigen. Clin Exp Metastasis. 9:283–299. 1991. View Article : Google Scholar : PubMed/NCBI
33	Bettenworth D, Mucke MM, Schwegmann K, Faust A, Poremba C, Schäfers M, Domagk D and Lenz P: Endoscopy-guided orthotopic implantation of colorectal cancer cells results in metastatic colorectal cancer in mice. Clin Exp Metastasis. 33:551–562. 2016. View Article : Google Scholar : PubMed/NCBI
34	Zawadzki A, Liu Q, Wang Y, Melander A, Jeppsson B and Thorlacius H: Verapamil inhibits L-type calcium channel mediated apoptosis in human colon cancer cells. Dis Colon Rectum. 51:1696–1702. 2008. View Article : Google Scholar : PubMed/NCBI
35	Wu Y, Guo Z, Zhang D, Zhang W, Yan Q, Shi X, Zhang M, Zhao Y, Zhang Y, Jiang B, et al: A novel colon cancer gene therapy using rAAVmediated expression of human shRNA-FHL2. Int J Oncol. 43:1618–1626. 2013. View Article : Google Scholar : PubMed/NCBI
36	Wang J, Yang Y, Xia HH, Gu Q, Lin MC, Jiang B, Peng Y, Li G, An X, Zhang Y, et al: Suppression of FHL2 expression induces cell differentiation and inhibits gastric and colon carcinogenesis. Gastroenterology. 132:1066–1076. 2007. View Article : Google Scholar : PubMed/NCBI
37	Amann T, Egle Y, Bosserhoff AK and Hellerbrand C: FHL2 suppresses growth and differentiation of the colon cancer cell line HT-29. Oncol Rep. 23:1669–1674. 2010.PubMed/NCBI
38	Al-Haidari A, Algaber A, Madhi R, Syk I and Thorlacius H: miR-155-5p controls colon cancer cell migration via post-transcriptional regulation of Human Antigen R (HuR). Cancer Lett. 421:145–151. 2018. View Article : Google Scholar : PubMed/NCBI
39	Liu C, Kelnar K, Liu B, Chen X, Calhoun-Davis T, Li H, Patrawala L, Yan H, Jeter C, Honorio S, et al: The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44. Nat Med. 17:211–215. 2011. View Article : Google Scholar : PubMed/NCBI
40	Takeyama H, Yamamoto H, Yamashita S, Wu X, Takahashi H, Nishimura J, Haraguchi N, Miyake Y, Suzuki R, Murata K, et al: Decreased miR-340 expression in bone marrow is associated with liver metastasis of colorectal cancer. Mol Cancer Ther. 13:976–985. 2014. View Article : Google Scholar : PubMed/NCBI
41	Algaber A, Al-Haidari A, Madhi R, Rahman M, Syk I and Thorlacius H: MicroRNA-340-5p inhibits colon cancer cell migration via targeting of RhoA. Sci Rep. 10:169342020. View Article : Google Scholar : PubMed/NCBI
42	Yang L, Men WL, Yan KM, Tie J, Nie YZ and Xiao HJ: miR-340-5p is a potential prognostic indicator of colorectal cancer and modulates ANXA3. Eur Rev Med Pharmacol Sci. 22:4837–4845. 2018.PubMed/NCBI
43	Wu ZS, Wu Q, Wang CQ, Wang XN, Huang J, Zhao JJ, Mao SS, Zhang GH, Xu XC and Zhang N: miR-340 inhibition of breast cancer cell migration and invasion through targeting of oncoprotein c-Met. Cancer. 117:2842–2852. 2011. View Article : Google Scholar : PubMed/NCBI
44	Xiao H, Yu L, Li F, Wang H, Li W and He X: miR-340 suppresses the metastasis by targeting EphA3 in cervical cancer. Cell Biol Int. 42:1115–1123. 2018. View Article : Google Scholar : PubMed/NCBI
45	Fernandez S, Risolino M, Mandia N, Talotta F, Soini Y, Incoronato M, Condorelli G, Banfi S and Verde P: miR-340 inhibits tumor cell proliferation and induces apoptosis by targeting multiple negative regulators of p27 in non-small cell lung cancer. Oncogene. 34:3240–3250. 2015. View Article : Google Scholar : PubMed/NCBI
46	Wu M, Wang J, Tang W, Zhan X, Li Y, Peng Y, Huang X, Bai Y, Zhao J, Li A, et al: FOXK1 interaction with FHL2 promotes proliferation, invasion and metastasis in colorectal cancer. Oncogenesis. 5:e2712016. View Article : Google Scholar : PubMed/NCBI
47	Park SY, Lee SJ, Cho HJ, Kim TW, Kim JT, Kim JW, Lee CH, Kim BY, Yeom YI, Lim JS, et al: Dehydropeptidase 1 promotes metastasis through regulation of E-cadherin expression in colon cancer. Oncotarget. 7:9501–9512. 2016. View Article : Google Scholar : PubMed/NCBI
48	Bai S, Zeng R, Zhou Q, Liao W, Zhang Y, Xu C, Han M, Pei G, Liu L, Liu X, et al: Cdc42-interacting protein-4 promotes TGF-B1-induced epithelial-mesenchymal transition and extracellular matrix deposition in renal proximal tubular epithelial cells. Int J Biol Sci. 8:859–869. 2012. View Article : Google Scholar : PubMed/NCBI
49	Yue B, Qiu S, Zhao S, Liu C, Zhang D, Yu F, Peng Z and Yan D: LncRNA-ATB mediated E-cadherin repression promotes the progression of colon cancer and predicts poor prognosis. J Gastroenterol Hepatol. 31:595–603. 2016. View Article : Google Scholar : PubMed/NCBI
50	Wendt MK, Taylor MA, Schiemann BJ and Schiemann WP: Down-regulation of epithelial cadherin is required to initiate metastatic outgrowth of breast cancer. Mol Biol Cell. 22:2423–2435. 2011. View Article : Google Scholar : PubMed/NCBI
51	Fan L, Wang H, Xia X, Rao Y, Ma X, Ma D, Wu P and Chen G: Loss of E-cadherin promotes prostate cancer metastasis via upregulation of metastasis-associated gene 1 expression. Oncol Lett. 4:1225–1233. 2012. View Article : Google Scholar : PubMed/NCBI